[The complications of laser surgery for the treatment of chronic vasomotor rhinitis].

The objective of the present study was to reveal cases of complications developing after laser coagulation of inferior turbinated bones. This treatment was given to 322 patients presenting with vasomotor rhinitis. A surgical ND:YAG laser with a quartz light guide 400 mcm in diameter was used for the purpose. In the majority of the cases, a single irradiation session produced a persistent positive effect, both functional and morphological. At the same time, two patients developed an unusual complication in the form of osteomyelitis of an inferior turbinated bone with its sequestration.

Search for virus nucleic acid sequences in postmortem human brain tissue using in situ hybridization technology with cloned probes: some solutions and results on progressive multifocal leukoencephalopathy and subacute sclerosing panencephalitis tissue.

In order to obtain a useful and readily applicable in situ hybridization (ISH) protocol for progressive central nervous system (CNS) diseases of unknown etiology that are possibly due to persistent viral infection, known and well described diseases were studied, namely, progressive multifocal leukoencephalopathy (PML) and subacute sclerosing panencephalitis (SSPE). The procedures described were validated by confirming results obtained by other investigators using histology, immunocytochemistry, electron microscopy, and ISH. A number of frequently encountered problems of tissue preparation are addressed as well as techniques to reduce autoradiography exposure times. A multi-staged specific, sensitive, reliable, and valid procedure for detection of viral genomes, mRNA and proteins is approached. Formalin-fixed and paraffin-embedded (FFPE) brain material from six patients who died with PML and one patient who died from SSPE were studied using ISH with a tritium-labeled cloned JC virus DNA probe and a measles-cloned nucleocapsid (NC) gene cDNA probe, respectively. This report constitutes a methodological framework as well as a detailed neuropathological analysis of identified brain cell populations within which in situ hybridization was detected. In early PML lesions, swollen nuclei or oligodendrocytes were the predominant cells labeled, whereas older lesions revealed increased numbers of reactive and bizarre hypertrophic astrocytes hybridized at the outer periphery of the demyelinated lesions. The hybridization varied greatly in intensity in different cells. Intense hybridization was noted very rarely in microglial cells, including rod cells and rarely in venular pericytes, intravascular mononuclear cells, or in vascular endothelial cells. These results, considered together with previous findings, indicate that in PML the viral infection runs different courses in the various cells: in astrocytes the viral genome persists for a long time inducing pathological changes in some cells. In oligodendrocytes the infection rapidly lyses the cells. There was a good correlation between chromatic changes observable in routinely stained sections and virus presence. In addition, in situ hybridization using a measles-NP-cloned probe in white matter from FFPE SSPE brain is presented confirming earlier results in SSPE cryopreserved brain.

Subacute sclerosing panencephalitis: measles virus matrix protein nucleic acid sequences detected by in situ hybridization.

Subacute sclerosing panencephalitis (SSPE) is characterized by a hyperimmune state toward the polypeptides of measles virus except the matrix (M) protein. Using cloned (3H)-labeled complementary DNA probes for in situ hybridization, we found the M protein and nucleocapsid (NP) protein nucleotide sequences in glial cells and neurons of cryostat sections from two SSPE brains. In one SSPE brain, M protein was lacking, but the other measles polypeptides were present. IgG and IgM antibodies eluted from that brain lacked antibodies to M protein, but antibodies to other measles polypeptides were present. In SSPE brain, the viral M-protein defect is not a deletion of the M gene, but rather a block in gene expression.

[Patient with progressive multifocal leukoencephalopathy by in situ hybridization: comparison of in situ hybridization using isotopic and biotinylated probes].

Ward et al developed biotinylated probes to hasten virus nucleic acid detection. We used these probes to trace SV 40 and JC virus nucleic acids in PML brain. This procedure visualizes the hybridization of viral sequences by affinity cytochemistry with avidin-biotin peroxidase complexes and diaminobenzidine. In the PML frozen white matter numerous oligodendroglial nuclei hybridized with JC virus biotinylated and tritium labelled probes in areas adjacent to active demyelination. Although tritium labeled probes was most sensitive and gave lowest background this biotinylated method showed be useful as a rapid diagnostic test for the specific detection of viral nucleic acid sequences in brain tissues from patients with central nervous system (CNS) infectious diseases.